Site-specific DNA methylation by a complex of PU.1 and Dnmt3a/b
Site-specific DNA methylation by a complex of PU.1 and Dnmt3a/b
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DOI：
10.1038/sj.onc.1209272
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发表时间：
2006-04
期刊：
影响因子：
8
通讯作者：
Mitsuhiro Suzuki;Mitsuhiro Suzuki;Toshiyuki Yamada;F. Kihara-Negishi;Takuya Sakurai;E. Hara;D. Tenen;N. Hozumi;T. Oikawa
中科院分区：
文献类型：
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作者：
Mitsuhiro Suzuki;Mitsuhiro Suzuki;Toshiyuki Yamada;F. Kihara-Negishi;Takuya Sakurai;E. Hara;D. Tenen;N. Hozumi;T. Oikawa
Ets转录因子PU. 1是骨髓和B细胞谱系发育所必需的造血主调节因子。如前所述，PU。1有时会抑制转录，与mSin3A-组蛋白脱乙酰转移酶-MeCP2形成复合物。在这里，我们展示了PU之间的相互作用。1和DNA甲基转移酶，DNA甲基转移酶（Dnmt）3a和Dnmt3b（Dnmt3s）。谷胱甘肽-S-转移酶下拉试验显示PU. 1通过ETS结构域直接与Dnmt3s的ATRX结构域相互作用。Dnmt3s抑制PU的转录活性。1在具有三聚化PU的报告子构建体上。1-结合位点。通过添加DNA甲基转移酶抑制剂5-氮杂脱氧胞苷而不是组蛋白脱乙酰酶抑制剂阿司他丁A来恢复抑制。亚硫酸氢盐序列分析显示，在启动子区域中的几个CpG位点邻近PU。1-当Dnmt 3s与PU共表达时，结合位点被甲基化。1.我们还发现，在p16 INK4A启动子的CpG位点甲基化的PU过表达。1在NIH3T3细胞中的表达，伴随着p16 INK4A基因表达的下调。这些结果表明，PU。1可能通过表观遗传修饰如DNA甲基化下调其靶基因。
The Ets transcription factor PU. 1 is a hematopoietic master regulator essential for the development of myeloid and B-cell lineages. As we previously reported, PU. 1 sometimes represses transcription on forming a complex with mSin3A–histone deacetyl transferase–MeCP2. Here, we show an interaction between PU. 1 and DNA methyltransferases, DNA methyltransferase (Dnmt) 3a and Dnmt3b (Dnmt3s). Glutathione-S-transferase pulldown assay revealed that PU. 1 directly interacted with the ATRX domain of Dnmt3s through the ETS domain. Dnmt3s repressed the transcriptional activity of PU. 1 on a reporter construct with trimerized PU. 1-binding sites. The repression was recovered by addition of 5-aza-deoxycitidine, a DNA methyltransferase inhibitor, but not trichostatin A, a histone deacetylase inhibitor. Bisulfite sequence analysis revealed that several CpG sites in the promoter region neighboring the PU. 1-binding sites were methylated when Dnmt3s were coexpressed with PU. 1. We also showed that the CpG sites in the p16 INK4A promoter were methylated by overexpression of PU. 1 in NIH3T3 cells, accompanied by a downregulation of p16 INK4A gene expression. These results suggest that PU. 1 may downregulate its target genes through an epigenetic modification such as DNA methylation.